LOCUS       sequence-238424-        8170 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8170)
  AUTHORS   Scaffidi P, Misteli T
  TITLE     Lamin A-dependent misregulation of adult stem cells associated with 
            accelerated ageing.
  JOURNAL   Nat Cell Biol. 2008 Apr;10(4):452-9. doi: 10.1038/ncb1708. Epub 2008
            Mar 2.
  PUBMED    18311132
REFERENCE   2  (bases 1 to 8170)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1038/ncb1708"; 
            journalName: "Nat Cell Biol"; date: "2008-04"; volume: "10"; issue: 
            "4"; pages: "452-9"
COMMENT     Sequence Label: pBABE-puro-GFP-wt-lamin A
FEATURES             Location/Qualifiers
     source          1..8170
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             130..546
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     476..498
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     514..530
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      587..596
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(593..595,596..598,599..1309)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(593..595,596..598,599..1309)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(638..659)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(899..918)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1246..1267
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3688..3708)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3693..4022
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3873..4008
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4032..4631
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4032..4051)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4528..4548
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4852..5321
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5451..5468)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5622..6210)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5702..5721)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6370..7161,7162..7230))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6993..7012
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7231..7335)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             7366..7835
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    7900..8099
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 ttcccgcctc cgtctgaatt tttgctttcg gtttggaacc gaagccgcgc gtcttgtctg
       61 ctgcagcgct gcagcatcgt tctgtgttgt ctctgtctga ctgtgtttct gtatttgtct
      121 gaaaattagg gccagactgt taccactccc ttaagtttga ccttaggtca ctggaaagat
      181 gtcgagcgga tcgctcacaa ccagtcggta gatgtcaaga agagacgttg ggttaccttc
      241 tgctctgcag aatggccaac ctttaacgtc ggatggccgc gagacggcac ctttaaccga
      301 gacctcatca cccaggttaa gatcaaggtc ttttcacctg gcccgcatgg acacccagac
      361 caggtcccct acatcgtgac ctgggaagcc ttggcttttg acccccctcc ctgggtcaag
      421 ccctttgtac accctaagcc tccgcctcct cttcctccat ccgccccgtc tctccccctt
      481 gaacctcctc gttcgacccc gcctcgatcc tccctttatc cagccctcac tccttctcta
      541 ggcgccggcc ggatcccagt gtggtggtac gtaggcgcta ccggtcgcca ccatggtgag
      601 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
      661 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
      721 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
      781 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
      841 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
      901 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
      961 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1021 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     1081 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     1141 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1201 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1261 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt ccggactcag
     1321 atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcagcg ctgccaacct
     1381 gccggccatg gagaccccgt cccagcggcg cgccacccgc agcggggcgc aggccagctc
     1441 cactccgctg tcgcccaccc gcatcacccg gctgcaggag aaggaggacc tgcaggagct
     1501 caatgatcgc ttggcggtct acatcgaccg tgtgcgctcg ctggaaacgg agaacgcagg
     1561 gctgcgcctt cgcatcaccg agtctgaaga ggtggtcagc cgcgaggtgt ccggcatcaa
     1621 ggccgcctac gaggccgagc tcggggatgc ccgcaagacc cttgactcag tagccaagga
     1681 gcgcgcccgc ctgcagctgg agctgagcaa agtgcgtgag gagtttaagg agctgaaagc
     1741 gcgcaatacc aagaaggagg gtgacctgat agctgctcag gctcggctga aggacctgga
     1801 ggctctgctg aactccaagg aggccgcact gagcactgct ctcagtgaga agcgcacgct
     1861 ggagggcgag ctgcatgatc tgcggggcca ggtggccaag cttgaggcag ccctaggtga
     1921 ggccaagaag caacttcagg atgagatgct gcggcgggtg gatgctgaga acaggctgca
     1981 gaccatgaag gaggaactgg acttccagaa gaacatctac agtgaggagc tgcgtgagac
     2041 caagcgccgt catgagaccc gactggtgga gattgacaat gggaagcagc gtgagtttga
     2101 gagccggctg gcggatgcgc tgcaggaact gcgggcccag catgaggacc aggtggagca
     2161 gtataagaag gagctggaga agacttattc tgccaagctg gacaatgcca ggcagtctgc
     2221 tgagaggaac agcaacctgg tgggggctgc ccacgaggag ctgcagcagt cgcgcatccg
     2281 catcgacagc ctctctgccc agctcagcca gctccagaag cagctggcag ccaaggaggc
     2341 gaagcttcga gacctggagg actcactggc ccgtgagcgg gacaccagcc ggcggctgct
     2401 ggcggaaaag gagcgggaga tggccgagat gcgggcaagg atgcagcagc agctggacga
     2461 gtaccaggag cttctggaca tcaagctggc cctggacatg gagatccacg cctaccgcaa
     2521 gctcttggag ggcgaggagg agaggctacg cctgtccccc agccctacct cgcagcgcag
     2581 ccgtggccgt gcttcctctc actcatccca gacacagggt gggggcagcg tcaccaaaaa
     2641 gcgcaaactg gagtccactg agagccgcag cagcttctca cagcacgcac gcactagcgg
     2701 gcgcgtggcc gtggaggagg tggatgagga gggcaagttt gtccggctgc gcaacaagtc
     2761 caatgaggac cagtccatgg gcaattggca gatcaagcgc cagaatggag atgatccctt
     2821 gctgacttac cggttcccac caaagttcac cctgaaggct gggcaggtgg tgacgatctg
     2881 ggctgcagga gctggggcca cccacagccc ccctaccgac ctggtgtgga aggcacagaa
     2941 cacctggggc tgcgggaaca gcctgcgtac ggctctcatc aactccactg gggaagaagt
     3001 ggccatgcgc aagctggtgc gctcagtgac tgtggttgag gacgacgagg atgaggatgg
     3061 agatgacctg ctccatcacc accacggctc ccactgcagc agctcggggg accccgctga
     3121 gtacaacctg cgctcgcgca ccgtgctgtg cgggacctgc gggcagcctg ccgacaaggc
     3181 atctgccagc ggctcaggag cccaggtggg cggacccatc tcctctggct cttctgcctc
     3241 cagtgtcacg gtcactcgca gctaccgcag tgtggggggc agtgggggtg gcagcttcgg
     3301 ggacaatctg gtcacccgct cctacctcct gggcaactcc agcccccgaa cccagagccc
     3361 ccagaactgc agcatcatgt aatctgggac ctgccaggca ggggtggggg tggaggcttc
     3421 ctgcgtcctc ctcacctcat gcccaccccc tgccctgcac gtcatgggag ggggcttgaa
     3481 gccaaagaaa aataaccctt tggttttttt cttctgtatt tttttttcta agagaagtta
     3541 ttttctacag tggttttata ctraaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     3601 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa agcgtctaga aaccctttag atctgctgca
     3661 gcgcgccagc acagtggtcg accctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3721 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3781 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3841 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3901 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3961 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4021 aaaagcttac catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
     4081 ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
     4141 tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
     4201 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
     4261 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
     4321 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
     4381 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca
     4441 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc
     4501 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
     4561 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
     4621 ccggtgcctg acgcccgccc cacgacccgc agcgcccgac cgaaaggagc gcacgacccc
     4681 atgcatcgat aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc
     4741 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata
     4801 cataactgag aatagagaag ttcagatcaa ggtcaggaac agatggaaca gctgaatatg
     4861 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
     4921 gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
     4981 ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc
     5041 agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca
     5101 atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc
     5161 cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt
     5221 atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt
     5281 ctcctctgag tgattgacta cccgtcagcg ggggtctttc acatgcagca tgtatcaaaa
     5341 ttaatttggt tttttttctt aagtatttac attaaatggc catagttgca ttaatgaatc
     5401 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5461 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5521 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5581 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5641 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5701 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5761 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5821 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5881 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5941 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6001 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6061 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6121 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6181 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6241 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6301 atcttcacct agatcctttt gcggccgcaa atcaatctaa agtatatatg agtaaacttg
     6361 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6421 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6481 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6541 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6601 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6661 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6721 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6781 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6841 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6901 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6961 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7021 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7081 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7141 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7201 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7261 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7321 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gcagcctgaa tatgggccaa
     7381 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag
     7441 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     7501 gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt
     7561 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
     7621 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
     7681 ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa
     7741 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
     7801 tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga
     7861 gacccctgcc cagggaccac cgacccacca ccgggaggca agctggccag caacttatct
     7921 gtgtctgtcc gattgtctag tgtctatgac tgattttatg cgcctgcgtc ggtactagtt
     7981 agctaactag ctctgtatct ggcggacccg tggtggaact gacgagttct gaacacccgg
     8041 ccgcaaccct gggagacgtc ccagggactt tgggggccgt ttttgtggcc cgacctgagg
     8101 aagggagtcg atgtggaatc cgaccccgtc aggatatgtg gttctggtag gagacgagaa
     8161 cctaaaacag
//
